A case of microphthalmos with cyst and partial trisomy 22.

A 13-day-old girl was referred for evaluation of a right orbital mass. Ophthalmic examinations revealed microphthalmia with cyst in the right eye and microphthalmia in the left eye. She had lowset ears, auricular fistula, micrognathia, and muscular hypotonia. Chromosomal analysis using fluorescence in-situ hybridization (FISH) showed partial trisomy 22. To our knowledge, microphthalmos with cyst associated with partial trisomy 22 has not been described previously in the English literature. Repeated aspiration resulted in a decrease in size of the cyst.

Calbindin-D 28 kD and parvalbumin in the horizontal cells of rat retina during development.

PURPOSE: Calbindin-D 28K (CaD) and parvalbumin (PV) are calcium-binding proteins thought to act as an intraneural calcium ion buffering system in the central nervous system. We previously reported that PV appears at birth in some cells in the outer portion of the nuclear layer of the retina that can differentiate into horizontal cells. CaD is also distributed in horizontal cells; however, it is not clear at which developmental stage CaD appears. METHODS: The development of horizontal cells and the distribution of CaD and PV was examined ultrastructurally and immunohistochemically in rat retinas from birth (P0) to postnatal day 50 (P50). RESULTS: Our ultrastructural data showed clear differentiation of horizontal cells from other retinal cells in the early postnatal days. These cells became nearly mature around P10 to P12. PV appeared in the outer nuclear layer at birth (P0) and, by P7, was observed in the ganglion cell layer, amacrine cells, and horizontal cells. CaD-immunoreactive cells, however, were not seen in any portions of the retina until P7. At P10, CaD-immunoreactivity was faintly observed in horizontal cells, and its immunoreactivity gradually increased in the horizontal cells from P10 to P15 when the eyes of the rats opened. CaD was not found in the ganglion cell layer or in amacrine cells at any period examined. CONCLUSION: In the horizontal cells, PV appeared when they were ultrastructurally distinguished from other retinal cells, while CaD appeared when they already had some structure for endocytosis and synaptic transmission, suggesting that PV and CaD may be good morphological and functional markers for horizontal cells, respectively.

Does light stimulus at eye opening of the developing rat influence retinal expression of GTP-binding protein (G(0))?

We previously hypothesized that light stimulus at eye opening of rats on postnatal days (P) 13 or 14 has an effect on the expression of GTP-binding proteins (G(0) in the retina) because the concentration of G(0) alpha increased rapidly between P10 and P15. This hypothesis was also supported by the findings that the distribution of G(0) alpha in the retina was almost the same as that of adult rats between P10 and P15. In this study, pregnant rats were kept in a dark room after vaginal plugs were identified; they gave birth to their pups in the dark, and their pups were reared by their mothers in the dark. The postnatal rats were sacrificed at P10, P15, P18, P22, P24, P27, and P30. Their retinas were investigated immunochemically and immunohistochemically, using G(0) alpha antibody, and these results were compared with those of the rat pups reared normally. Only G(0) alpha immunoreactivity in the inner nuclear layer of rats reared in the dark room was weaker than in the controls; the distribution of G(0) alpha in the retina did not change, as compared with pups reared in normal conditions. In addition, in pups reared in the dark, G(0) alpha increased rapidly from P10 to P15. However, the concentration of G(0) alpha in the retina of rat pups reared in the dark was significantly low at P22 (p < 0.01) and P30 (p < 0.05), as compared with pups reared in normal conditions. Although the function of G(0) in the retina may have something to do with the light stimulus after eye opening, it appeared that the expression of G(0) was not influenced by light stimulus at eye opening.

Choroidal osteoma in an infant.

PURPOSE: To report an 8-month-old infant with choroidal osteoma. METHOD: Case report. RESULTS: The patient, who had bilateral yellow stippling at the posterior pole, was followed up for 8 years. Both fundi developed creamy, irregular scalloped lesions. Computed tomography showed a bony plate at the posterior pole bilaterally. CONCLUSION: We believe that this is the youngest patient reported to have choroidal osteoma.

Comparison of immunostainings for proliferating cell nuclear antigen and Ki-67 in human extraocular lesions.

BACKGROUND: Cell kinetic information is helpful to understand disease progression, treatment response, and prognosis of the neoplasms. To compare the usefulness and limitations of antibodies that recognize the cell cycle-associated molecules, proliferating cell nuclear antigen (PCNA) and Ki-67, we performed immunostaining in formalin-fixed, paraffin-embedded tissue sections of extraocular lesions. METHODS: Specimens were obtained from patients undergoing routine surgical procedures. Formalin-fixed, paraffin-embedded sections were stained for PCNA and Ki-67 using the monoclonal antibodies PC10 and MIB-1, respectively. Microwave oven heating for antigen retrieval was performed before immunostaining. RESULTS: In squamous cell carcinomas and basal cell carcinomas, PCNA immunostaining varied greatly. Basal cells of benign epithelial lesions showed moderate to weak PCNA staining. Strong PCNA immunoreactivity was demonstrated in foci of inflammation and germinal centers. Microwave processing enhanced the intensity of those PCNA immunostainings. As the PCNA immunostaining intensity had a great variability, the absolute numbers of PCNA-positive cells were hard to count in some tissues. High Ki-67 counts were observed in squamous cell carcinomas, foci of inflammatory cells, and germinal center cells. Basal cell carcinomas and benign lesions showed low Ki-67 counts. Every section showed clear nuclear staining in Ki-67 immunostaining. CONCLUSION: Careful consideration is required in the assessment of cell proliferation using PCNA. The immunostaining of Ki-67 may be more accurate than that of PCNA for evaluating cell proliferation in formalin-fixed, paraffin-embedded tissues.

Ontogeny of GTP-binding proteins, Gi and G(o), in rat retina.

The distribution and the levels of Gi1 (plus Gi3), Gi2, and G(o) in rat retina were studied immunohistochemically and immunochemically during development. At embryonic day (E) 15, Gi1 alpha/Gi3 alpha was observed in the inner layer of the neural retina, the future nerve fiber layer (NFL), while Gi2 alpha was observed both in the inner and outer layers of the neural retina. No immunoreactivity for G(o) alpha was observed. At E18, Gi1 alpha/Gi3 alpha and Gi2 alpha appeared in the inner plexiform layer (IPL), while G(o) alpha was faintly immunoreactive only in the NFL. At birth, Gi2 alpha/Gi3 alpha and G(o) alpha appeared in the ganglion cell layer. Gi2 alpha was intensely immunoreactive in the NFL and IPL. At postnatal day (P) 10, the inner portions of the retina, from the NFL to the outer plexiform layer, were immunoreactive to Gi1 alpha/Gi3 alpha, Gi2 alpha, and G(o) alpha. Gi1 alpha/Gi3 alpha and G(o) alpha were distributed characteristically in a laminated pattern in the IPL, but Gi2 alpha was present homogeneously in the IPL. At P12, Gi2 alpha appeared in the outer nuclear layer. As the postnatal days advanced, the laminated pattern of immunoreactivity to G(o) alpha in the IPL became diffuse, but immunoreactivity to Gi1 alpha/Gi3 alpha remained. The results of enzyme immunoassays showed that the concentration of G(o) alpha increased rapidly from P10 to P15 and reached almost the adult level at P20-P30, while Gi2 alpha decreased until P15 and was almost constant thereafter. These results showed that the distribution of Gi1 alpha/Gi3 alpha, Gi2 alpha, and G(o) alpha differs during development, suggesting that each G protein in the developing retina has a unique function.

Chronic retinal effects by ultraviolet irradiation, with special reference to superoxide dismutases.

Recently ultraviolet light (UV) reaching the Earth's surface has been gradually increasing in amounts by the destruction of the ozone layers. Large parts of UV are absorbed in the cornea and lens, and only a few amounts reached the retina; however, the effect on the retina is not fully elucidated. 38 rats were irradiated 0.5-5.0 J/cm2 UV from 6 to 50 times every 24 hours, and immunohistochemically and immunochemically for superoxide dismutases (SOD). Morphologically, the destruction of rod outer segments (ROS) and dissociation of cell membranes between the pigment epithelial cells (PE) were already observed by 6 times 0.5 J/cm2 UV irradiations. As the doses of UV increased, heterochromatins and lipid droplets increased in the PE. In normal retina, Cu/Zn SOD were mainly distributed from the inner limiting membrane (ILM) to the ganglion cell layer, and the PE; however, after 6 times 0.5 J/cm2 UV irradiations, the distribution became widened from inner to outer plexiform layer (OPL). At that time, the concentrations of Cu/Zn and Mn SOD increased in the retina. The present study reveals that the morphological damage caused by UV irradiation is observed in the ROS and PE, where no immunoreactivities could be detected to Cu/Zn and Mn SOD. However, morphological damage was not from the ILM to OPL, where the immunoreactivities to both Cu/Zn and Mn SOD were observed.

Effects of paraquat on mitochondrial electron transport system and catecholamine contents in rat brain.

The effects of paraquat on rat brain were studied. Activities of complex I (NADH: ubiquinone oxidoreductase) in mitochondrial electron transport system, lipid peroxidation and the amount of catecholamines in rat brain were measured after acute paraquat exposure. Complex I activities were significantly lower and lipid peroxides were higher in the brains of a paraquat-treated group than in those of a control group. Lipid peroxide in rat serum, however, did not increase after paraquat exposure. A study of the time dependency of paraquat effects disclosed that mitochondrial complex I activities in rat brain as well as those in rat lung and liver gradually decreased prior to the appearance of respiratory dysfunction. As compared to controls, the dopamine in rat striatum was significantly lower in the paraquat-treated group. These results suggest that paraquat after crossing the blood-brain barrier might be reduced to the radical in rat brain, which may damage the brain tissue, especially dopaminergic neurons in striatum. We therefore propose that cerebral damage should be taken into consideration on paraquat exposure. Patients may therefore need to be followed up after exposure to high doses of paraquat.

Immunohistochemical study of 28-kD heat shock protein in the lens of young human embryos.

The localization of 28-kD heat shock protein (HSP28) in the lenses of human embryos (Carnegie stages 13-21) was studied immunohistochemically using antibodies for HSP28. At stage 13, HSP28 was observed in the lens placode. At stage 15, when the lens vesicle is formed by the invagination of the lens placode, HSP28 was observed in the posterior portion of the lens vesicle. From stages 16 to 18, when the posterior portion of the lens epithelium elongates toward the anterior lens epithelium and forms the primary lens fiber, HSP28 was observed in the apical and basal sides of the primary lens fiber. At stage 19, when the primary lens fiber attaches to the anterior lens epithelium, immunoreactivity to HSP28 appeared in the anterior lens epithelium. At stage 21, the anterior lens epithelium and the equator portion of the lens, where the secondary lens fiber is formed, were intensely immunoreactive to HSP28. Although alpha B-crystallin is thought to be one of the small HSPs, the distribution of HSP28 in the lenses of human embryos was different from that of alpha B-crystallin which we previously reported.

Immunohistochemical localization of epidermal growth factor receptor and epithelial antigen in tumors of the human conjunctiva, eyelid, lacrimal gland, and orbit.

BACKGROUND: Increased numbers of epidermal growth factor (EGF) receptors are observed in squamous cell carcinomas of human lung, head, neck, and cervix. We studied the presence of EGF receptors and epithelial antigen in some ophthalmic lesions. METHODS: Immunohistochemical staining for EGF receptors was assessed in tumors of human conjunctiva, eyelid, lacrimal glands, and orbit with monoclonal antibodies (EGF-R1 and clone 29.1). Reactivity of Ber-EP4, which recognizes epithelial antigen, was also examined. RESULTS: Strong staining of EGF-R1 and clone 29.1 and weak to moderate staining of Ber-EP4 were demonstrated in conjunctival squamous cell carcinomas. Cell membranes of conjunctival papilloma were moderately or strongly stained with these antibodies. Ductal components in sebaceous gland adenoma of the eyelid and pleomorphic adenoma of the lacrimal gland were positively stained. The antibodies did not bind to reactive lymphoid hyperplasia of the orbit and Wegener's granulomatosis. Relatively good correlation for immunostaining reaction was observed among EGF-R1, clone 29.1, and Ber-EP4 in each tumor. CONCLUSION: Immunostaining using EGF-R1, clone 29.1, and Ber-EP4 may be useful in differentiating epithelial tumors from non-epithelial lesions. Strong immunostaining for EGF receptor may be the hallmark of epidermoid malignancy.

Pyrroloquinoline quinone (PQQ) inhibits the expression of tyrosinase mRNA by alpha-melanocyte stimulating hormone in murine B16 melanoma cells.

The biological functions of pyrroloquinoline quinone (PQQ), a bacterial redox coenzyme and potent radical scavenger, have not been elucidated in mammals. In this paper, we studied the effects of PQQ on tyrosinase activity and subsequent melanogenesis in murine B16-F10 melanoma and found that alpha-Melanocyte stimulating hormone (MSH)-induced melanogenesis was inhibited by 6.3 to 25 microM PQQ in a dose-dependent manner. Moreover, PQQ inhibited MSH-induced tyrosinase activity by suppressing tyrosinase mRNA expressed by MSH. However, PQQ had no effect on MSH-stimulated cyclic adenosine 3', 5'-monophosphate (cAMP) production. These observations suggest that PQQ inhibits the expression of tyrosinase mRNA at a post receptor level and that PQQ may be useful in investigating hormone actions mediated by cAMP.

Ontogeny of copper-zinc and manganese superoxide dismutase in the developing rat retina: immunohistochemical and immunochemical study.

The localization of copper-zinc and manganese superoxide dismutase (SOD) in the developing rat retina was studied immunohistochemically and immunochemically. Immunoreactivity to Cu/Zn SOD was observed in the inner limiting membrane, nerve fiber layer, ganglion cell layer (GCL) and pigment epithelium on postnatal day (P) 7. From P7 to at least 10 weeks, the distribution of Cu/Zn SOD remained unchanged in rats. From P15 to P30, the immunoreactivity to Mn SOD appeared in the GCL and inner plexiform layer. The distribution of Mn SOD remained unchanged between P30 and 10 weeks. Our immunochemical study revealed that the concentration of Cu/Zn SOD was higher than that of Mn SOD throughout the postnatal period. The differential distributions of Cu/Zn SOD in the developing rat retina support the hypothesis that Cu/Zn and Mn SOD play an important role in the protection against oxygen free radicals in the different layers of the retina throughout development.

The effects of PSK, a biological response modifier, on congenital ocular abnormalities induced by X-ray irradiation.

The antiteratogenic effects of PSK, a biological response modifier, were examined using histological and developmental analysis. The whole bodies of pregnant mice were irradiated with X-rays and injected with PSK within ten minutes after irradiation on day 7 of gestation (E7). The foetuses on E18 were examined and a high incidence of malformations were observed in X-ray irradiated embryos. Microphthalmia was the most frequent malformation. PSK administration suppressed the X-ray irradiation-induced ocular anomalies in not only the frequency, as deduced by external observation, but also in histopathological changes in the retina, lens, and cornea. In particular, the incidence of lens aplasia was significantly decreased by PSK administration. Developmental analysis using E10 and E13 embryos revealed that the decrease in the incidence of histopathological changes was first observed within 72 hours after PSK administration. In addition, X-ray irradiation-induced early foetal death (E10-13) was also suppressed by PSK administration. The possible mechanisms of the antiteratogenic effects of PSK are discussed.

Preventive effect of SA3443, a novel cyclic disulfide, on glucocorticoid-induced cataract formation of developing chick embryo.

The preventive effect of SA3443 [(4R)-hexahydro-7,7-dimethyl-6-oxo-1,2,5-dithiazocine-4-carboxylic acid] against glucocorticoid-induced cataract of developing chick embryos was studied. When hydrocortisone succinate sodium (HC: 0.25 mumol per egg) was administered to 15-day-old embryos, almost all lenses became opaque (stage I:O%, II: 2.5 +/- 4.6%, III: 5 +/- 5.4%, IV-V 92.5 +/- 7.1%) at 48 hr after the treatment. However, a double application of SA3443 (10 mumol per egg) at 3 and 10 hr after HC treatment effectively prevented the cataract formation (stage I: 52.8 +/- 13.7%, II: 11.6 +/- 6.3%, III: 22.9 +/- 8.9%, IV-V: 13.9 +/- 11.0%) and diminished the decline in glutathione in the lens at 48 hr and in the liver at 24 hr after HC administration. The cleavage of the cyclic disulfide bond of SA3443 did not occur in the lens homogenate but in the liver homogenate. These results suggest that the appearance of sulfhydryl residue in the liver may contribute to the anticataract effects by representing radical scavenger activities.

[Development of experimental model animals for disseminated Mycobacterium avium complex infections using immunodeficient mice and rats].

In order to establish an animal model for disseminated M. avium complex (MAC) infections frequently encountered in AIDS patients, we studied growth of M. intracellulare in visceral organs (lungs, livers, spleens, kidneys), in blood, and in footpads of mice with defined immunodeficiencies, such as SCID mice with T and B cell-defect, BALB/c athymic nude mice with matured T cell-defect, and beige mice with NK cell-defect. In addition, Sprague-Dawley rats with acquired immunodeficiency induced by cyclosporine-treatment were also examined. The following results were obtained. 1) SCID mice: First, SCID mice were infected sc with 6.1 x 10(6) CFU of M. intracellulare N-260 (virulent SmT colonial variant) into the hind footpad. The organisms grew in the footpad remarkably during the 12 weeks after infection in SCID mice, where the growth rate was much greater than that in CB-17 strain mice with the same genotype as SCID mice and in BALB/c mice with Bcgs genotype (CB-17 and BALB/c mice are MAC-susceptible). Furthermore, in SCID mice, bacteremia and dissemination of organisms to the visceral organs were observed but not in the two control strains of mice. Second, SCID mice were infected i.v. with 4.8 x 10(6) CFU. The bacterial loads in the viscera of SCID mice after infection were larger than those of CB-17 mice except for livers. However, the incidence and the degree of gross lung lesions were much less in SCID mice compared to CB-17 mice, presumably due to the defect in T cell-mediated immune reactions in SCID mice.(ABSTRACT TRUNCATED AT 250 WORDS)

Ontogeny of alpha-crystallin subunits in the lens of human and rat embryos.

The distribution of alpha A- and alpha B-crystallin in the developing lens of human (Carnegie stages 13 to 23) and rat embryos (embryonic days E11 to 18) was examined immunohistochemically. In a human embryo at stage 13, the lens placode was already immunoreactive to alpha B-crystallin, but not to alpha A-crystallin. At stage 15, the lens vesicle was intensely immunoreactive both to alpha A- and alpha B-crystallin. From stages 16 to 23, the lens epithelial cells and fiber cells were immunoreactive to alpha A- and alpha B-crystallin. In rat embryos, alpha A-crystallin appeared in the lens pit at E12, and alpha B-crystallin appeared in the elongating lens fiber cells at E14. From E15 to E18, the lens epithelial cells and fiber cells were immunoreactive to alpha A-crystallin. The lens fiber cells were also immunoreactive to alpha B-crystallin, but the epithelial cells were not. These findings suggest that alpha B-crystallin appears earlier than alpha A-crystallin in the human lens, but at a later period than alpha A-crystallin in the rat lens. alpha B-Crystallin was not detected in the epithelial cells of the rat lens, but was persistently present in the epithelial cells of the human lens.

High myopia, retinal dystrophy, and mental retardation in siblings.

A 15-year-old girl and her 13-year-old brother had high myopia, mottled retinas with nonrecordable electroretinographic responses OU, and mental retardation. Both siblings had normal pupillary reactions to light, normal levels of serum amino acids, no nystagmus, no obesity, and no polydactyly. A consanguineous marriage of the patients' parents was found. We believe that the association of high myopia, retinal dystrophy, and mental retardation noted in these siblings may be uncommon.

Development of the lens in human embryos: a histochemical and ultrastructural study.

The early development of the lens was examined, using 36 externally normal human embryos at Carnegie stages 13-23 (4 to 8 weeks of gestation). Twenty-two embryos were sectioned serially and stained with periodic acid-Schiff and a modified method of PAS. In 14 embryos, not only the differential distribution of glycogen but also the ultrastructural change in the developing lens, with special reference to junctional complexes, were examined electron microscopically. At stage 15, when the lens vesicle was formed, glycogen was observed in the cytoplasm of the lens epithelium, especially in the posterior lens epithelium. From stages 16 to 18, when the posterior lens epithelium was differentiated into the primary lens fibers and elongated toward the anterior lens epithelium, the amount of glycogen increased in the basal cytoplasm of the primary lens fiber, where the intracellular organelles, such as the tubular vesicles, mitochondria and multivesicular bodies, began to aggregate. At stage 20, when the lens cavity was obliterated, glycogen was also present in the anterior lens epithelium. At stage 21, as the formation of the secondary lens fibers proceeded, glycogen was noted in the secondary lens fibers in the equator region. These findings suggest that the distribution of glycogen is associated with the formation of the primary and secondary lens fibers. In addition, we provide additional information that a lot of glycogen is distributed in the region where many intracellular organelles aggregate in the embryonic lens vesicles.

Viscoelastic substance in the anterior chamber elevates intraocular pressure.

An 85-year-old man underwent an extracapsular cataract extraction OD with posterior chamber intraocular lens implantation. Sodium hyaluronate (0.3 mL) was used during the procedure, and approximately 1.0 mL of the solution, including the sodium hyaluronate, was aspirated before wound closure. The next day, the intraocular pressure OD was elevated to 60 mmHg, and it remained high despite medication. Three days later, the intraocular pressure was still high, a paracentesis was done, and viscous solution was obtained. After the paracentesis, the intraocular pressure OD normalized.

Retrocorneal ridges after anterior posterior radial keratotomy (Sato's operation) in a patient with retinitis pigmentosa.

A 51-year-old man with bilateral cataracts and retinitis pigmentosa had decreased visual acuity. He also had retrocorneal ridges OU. The patient had undergone anterior posterior radial keratotomy (Sato's operation) OU at age 14 years. We believe that the retrocorneal ridges in our patient, which were found after Sato's operation, may be rare.